Sentences with phrase «of kahweol»

B) In situ determination of kahweol effects on HT - 1080 gelatinases, as determined by gelatin zymography with the presence of kahweol in the incubation substrate buffer.

All these effects may open a window for the potential therapeutical application of kahweol as an anti-angiogenic drug.

Although 50 nmol of kahweol per CAM was required to observe in vivo inhibition of angiogenesis in 100 % of treated eggs, as little as 10 nmol of kahweol was enough to induce clear inhibition of angiogenesis in 25 % of the tested eggs in the CAM assay.

Subconfluent HUVEC cultures were stimulated with PMA (50 ng / mL) for 4 h in the absence (controls) or presence of different concentrations of kahweol.

A third line of evidence showing the potential of kahweol to inhibit overall angiogenesis is provided by the ex vivo model of the mouse aortic ring assay.

The gelatinolytic assays were carried out in two different ways to obtain complementary information: firstly, cells were treated or not with the test compound and samples from these were submitted to gelatinase zymography to detect the effects of the kahweol treatment on the expression of gelatinase activities; secondly, in some experiments, samples from control, untreated HT - 1080 fibrosarcoma cells were submitted to zymography and, after electrophoresis, different concentrations of kahweol were added to the substrate buffer to determine the potential direct effect of kahweol on gelatinase activity.

Data obtained on the effects of kahweol on endothelial cell invasion (as determined by a continuous fluorescent assay) clearly show that kahweol induces an anti-invasive effect in HUVEC in a dose - dependent manner (Figure 7).

C) Quantification of the amount of MCP - 1 secreted by HUVEC after a 24 h treatment in the presence of different concentrations of kahweol.

We also demonstrate the inhibitory effect of kahweol on the endothelial cell potential to remodel extracellular matrix by targeting two key molecules involved in the process, MMP - 2 and uPA.

On the other hand, these data suggest that the potential effects of kahweol on apoptosis could exhibit certain cell specificity.

Furthermore, the inhibitory effects of kahweol on COX - 2 and MCP - 1 reinforce the idea of kahweol being a multi-targeted natural compound with high pharmacological potential.

Different in vitro assays were carried out in order to test the specific effects of kahweol treatment on several key steps of the angiogenic process in both endothelial and tumor cells.

Since invasion is dependent on extracellular matrix remodeling capabilities, this inhibitory effect strongly suggested that the two key extracellular membrane remodeling enzymes expressed by endothelial cells, namely, MMP - 2 and uPA could be other main key targets of the pharmacological action of kahweol on endothelial cells.

On the other hand, the present research work shows a confirmatory evidence of the potential of kahweol to inhibit in vivo angiogenesis, by using another completely independent model system, namely, that of genetically modified zebrafish.

Therefore, we studied the effects of kahweol on the growth of endothelial cells.

Therefore, it would be advisable to test the potential effects of kahweol on endothelial cell apoptosis.

These data may contribute to the explanation of the reported antitumoral effects of kahweol, including the recent epidemiological meta - analysis showing that drinking coffee could decrease the risk of certain cancers.

Our results reinforce the potential pharmacological interest of kahweol, as suggested by its anti-angiogenic and anti-inflammatory effects.

Six eggs were used for each tested dose of kahweol.

On the other hand, the anti-oxidant nature of kahweol also points to its potential anti-inflammatory capabilities.

To get new, additional insights on the features of kahweol as an anti-angiogenic compound, we carried out a complete set of in vitro assays previously used by us to characterize the anti-angiogenic effects of other compounds from natural sources, including aeroplysinin - 1, homocysteine, ursolic acid, puupehenone, hypericin, hyperforin and aloe - emodin, among others [11], [12], [13], [19], [20], [21].

Not exact matches

Furthermore, a clear dose - response inhibition of microvessel formation was observed and quantified in treatments with kahweol.

Figure 6 shows the effects of 75 µM kahweol on endothelial cell migration, as determined by the «wound healing» assay, after 8 and 24 h of treatment.

A) Gelatin zymography of MMP - 2 in conditioned media of HUVEC after treatment with different kahweol concentrations.

In the CAM assay, the inhibitory doses exhibited by kahweol are similar to those of other anti-angiogenic compounds found by us to inhibit angiogenesis in the CAM assay [10], [19], and much lower than those of other anti-angiogenic compounds [21].

The global morphological features (including centrifugal growth of the peripheral vessels - relative to the position of the disc -, avoiding the treated area, with an overall decrease in the vascular density) elicited by kahweol treatment are also in agreement with those previously observed for other anti-angiogenic compounds.

In all these in vitro assays, kahweol treatments were carried out under conditions (kahweol concentration and duration of treatment) that did produce no cytotoxic effect on cells.

The minimal inhibitory concentration for kahweol in this assay of «tubule - like» structures formation on Matrigel was 25 µM, in the range of concentrations at which other known antiangiogenic compounds produce this kind of effect [13], [27].

Figure 8 (C) shows that kahweol - treatment induces a dose - dependent decrease in the levels of urokinase in HUVEC conditioned media, with an almost complete inhibition at 50 µM kahweol.

The CAM and zebrafish in vivo assays and the ex vivo mouse aortic ring assay clearly identify kahweol as a new anti-angiogenic compound, but gives no information on which specific steps of angiogenesis are targeted by this compound.

The kahweol concentrations required to inhibit the differentiation of HUVEC cells did not affect their viability (results not shown).

Figure 2 (middle line) shows that 50 nmol kahweol induced apoptosis in a small percentage of cells but this effect did not seem to be endothelial cell specific, since only few apoptotic nuclei corresponded to endothelial cells.

One of these is kahweol (Figure 1), an antioxidant diterpene that remains in unfiltered coffee beverages, such as Turkish and Scandinavian coffee [3].

Video images of blood flow thru intersegmental vessels were taken on the caudal region next to vitellus in 48 h larvae after 24 h of treatment in the absence (control, Video S1) or presence (Video S2) of 50 µM kahweol.

Figure 9 (A and B) shows that kahweol inhibits in a dose dependent manner the expression of COX - 2 protein by HUVEC.

Survival curves of proliferative (squares) and non-proliferative (circles) HUVEC endothelial cells treated with kahweol.

Therefore, kahweol - treatment has another key target in this essential step of the angiogenesis process.

A) Negative of photographs (x20) of aortic rings (lateral view) after 10 days of incubation in a 3D collagen gel overlayed with complete medium in the presence of 20 mg / mL VEGF, 0.05 % DMSO (the vehicle taken as a control), or kahweol at 1, 5 and 25 µM (K1, K5, K25, respectively).

This effect on cell survival was not endothelial cell - specific, since IC50 values for kahweol treatment of several human tumoral cell lines were similar to those obtained for HUVEC (results not shown).

C) Plasminogen zymography of HUVEC uPA after treatment with different kahweol concentrations.

We show for the first time that kahweol is an anti-angiogenic compound with inhibitory effects in two in vivo and one ex vivo angiogenesis models, with effects on specific steps of the angiogenic process: endothelial cell proliferation, migration, invasion and tube formation on Matrigel.

Our results in the zymographic assays for gelatinase and urokinase activities clearly showed that, in fact, kahweol was able to inhibit the expression of both MMP - 2 and uPA, identifying them as two relevant molecular targets for kahweol.

It is believed that kahweol and cafestol palmitate increase the liver's production of glutathione, the master antioxidant, by as much as 700 %.

Research on the antioxidant activity of trigonelline, cafestol and kahweol has been less extensively investigated in humans.